Localization and therapy of non-prostatic endocrine cancer with agents directed against prostate specific antigen

ABSTRACT

It was discovered that prostate-specific antigen is produced by non-prostatic endocrine cancers. It was further discovered that non-prostatic endocrine cancers with steroid receptors can be stimulated with steroids to cause them to produce PSA either initially or at increased levels. This invention relates to the imaging of non-prostatic endocrine cancers by labelled biological binding units which bind to prostate-specific antigen in an imaging procedure, such as, radioimaging or magnetic resonance imaging. Further, the PSA-binding units may be constructed to deliver a toxic agent, such as a radioisotope, toxin or a drug to provide endocrine cancer therapy. Another aspect of the invention is passive immunotherapy against endocrine cancers by treatment with PSA-binding units.

FIELD OF THE INVENTION

This invention relates to the localization and therapy of non-prostatic endocrine cancers by agents that have been constructed to target prostate specific antigen (PSA).

BACKGROUND OF THE INVENTION

Cancer of the breast is the most common cause of cancer death in middle aged women in Europe and North America and both its incidence and mortality are on the increase (1-5). The predominant indications for breast tumor imaging are: detecting the presence of tumor, localizing sites of disease, and following the effects of therapy (6). Trends in scintigraphic imaging have been towards developing imaging pharmaceuticals to provide quantitative information on the pathophysiological characteristics of a tumor, such as its anaplasticity, or likely response to a given therapy (7). For example, to determine via scintigraphic imaging how a patients breast cancer will respond to the administration of a growth suppressor, such as somatostatin, or the estrogen receptor antagonist tamoxifen (8). Diamandis in PCT Application CA 94/00267 has shown that the presence of PSA in breast tumors, as measured by in vitro methods, has prognostic value. Thus, imaging of these tumors may not only reveal occult disease, but may also provide clinically valuable pathophysiological information.

Tumor imaging is commonly carried out using a gamma emitting radionuclide conjugate and a scintillation gamma camera, or with a positron emitting radiopharmaceutical and a positron or PET camera, or with a magnetopharmaceutical and a magnetic resonance imaging device. The scintillation camera, also known as an Anger camera, consists of a detector head, and a display console. The Anger camera head is composed of sodium iodide crystals that absorb gamma rays and emits the absorbed energy as flashes of light--scintillations that are proportional in brightness to the energy absorbed. In a gamma camera the sodium iodide crystals are coupled to photomultiplier tubes that convert light pulses into electronic pulses. These voltages are translated via a computing circuit to a cathode ray tube. The data from the camera head may be in either analog or digital form that can be stored in a computer and can reconstruct the data to provide an image. Single-photon emission computed tomography (SPECT) imaging involves the use of a gamma scintillation camera where multiple images, typically encompassing 180° or 360°, around the body are taken and the computer issued to reconstruct multiple tomograms in coronal, sagittal, and transverse projections. In PET imaging the positron radionuclide collides with an electron causing annihilation of the particles and creating two photons that travel in 180° opposite directions. The PET system is designed to capture opposite sides and register the count at precisely the same time. A computer is used to manipulate the data and then reconstruct a cross sectional image from this information.

There are a number of approaches to breast tumor imaging that may be divided into two groups: indirect and direct. Indirect techniques, are generally utilized to locate metastatic disease by recognizing the secondary effects of tumor within an organ system. Indirect techniques include, but are not limited to, the use of radiolabelled ^(99m) Tc phosphonates to locate bone metastases (9, 10) and ^(99m) Tc radiocolloids in liver scans and breast lyphoscintigraphy (11, 12, 13).

Direct approaches to radionuclide imaging include radiolabelled chemotherapeutic agents, simple ionic substances, metabolite imaging, immunologic and receptor imaging. The use of radiolabelled chemotherapeutic agents, such as bleomycin, have not demonstrated clinical value (14). ⁶⁷ Ga citrate is the most commonly used simple ionic tracer for tumor imaging, however it localizes in other pathologies and is non specific (15, 16, 17, 18). Metabolite imaging carried out with positron emitting radionuclides such as ¹⁸ F-fluorodeoxyglucose, ¹¹ C-methionine and ¹¹ C-thymidine provides tumor metabolism information that has been shown to be clinically valuable for disease staging (19, 20, 21).

The receptor imaging of breast cancer has been attempted by several approaches. Spicer et al. (22) and Hochberg (23) and others (24-29) have developed radiolabelled estradiols and have been able to demonstrate imaging in estrogen receptor positive breast cancers. It has been postulated that a therapeutic response could result with Auger electrons from ¹²³ I or ¹²⁵ I radiolabelled estradiols (30, 31), or from β emitting radioisotopes such as ¹⁸⁶ Re conjugated to progesterone (32). A problem with receptor based imaging is the interference that estrogen receptor antagonists, such as tamoxifen, may have in the clinical environment.

It is known that proteins, such as antibodies, can be developed against specific antigens that are either produced or associated with tumors, can be used to localize tumors. U.S. Pat. No. 3,927,193 to Hansen et al. (33) discloses a process whereby antibodies to carcinoembryonic antigen (CEA) and labelled with ¹²⁵ I and ¹³¹ I were used to image the location of tumors present in hamsters. From this work it was proposed that the location of a tumor in a human could be determined by in vivo administration of a parenteral solution containing an antibody-radioisotope conjugate followed by imaging by a gamma camera. Goldenberg et al. reported success in clinical trials of tumor detection and localization by scintillation scanning of patients that received radiolabelled antibodies to CEA (34).

Based on the original work of Milstein and Kohler (35), monoclonal antibodies have been developed against a variety of tumour antigens such as CA 19.9, CA 125, melanoma associated antigens, TAG 72, α fetal protein, ferritin, choriogonadotropin, prostatic acid phosphatase, and PSA for radioimmunoimaging and therapy.

Several investigators have reported on the development of monoclonal antibodies against epitopes of various malignant prostate cell components (36, 37, 38, 39, 40). Moveover, PSA was purified and well characterized and found to have a molecular weight in the range of 34,000 (41). PSA is used widely as a tumor marker for in vitro based analyses for diagnostic and monitoring purposes of prostatic carcinoma. U.S. Pat. No. 5,162,504 describes monoclonal antibodies that have been developed to recognize malignant prostate epithelium. These antibodies were developed as diagnostic and prognostic tools for the detection of cancer of the prostate, not as embodies in this invention, for the detection of cancers not associated with the prostate. Until the discovery reported by Diamandis in International Application PCT CA94/00267, it was thought that PSA only occurred in men and was only produced by prostate tissue.

To image breast tumors researchers have developed antibodies directed against TAG 72, CA-3, CEA, EGF-R, LASA-P, and other glycoproteins associated with breast cancer (42, 43, 44). Khaw et al. developed the monoclonal antibody 323/A3 against a 43 Kd membrane associated glycoprotein from the MCF-7 tumor cell line that was able to image tumors as small as 0.19 grams (45). Rainsbury et al. developed the antibody LICRCON-M8 against human milk fat globule and were also able to demonstrate imaging of human breast tumors, of particular note metastases to the bone were found (46).

Antibodies have been labelled directly with radioisotopes such as ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸ F, ¹⁸⁶ Re, ¹⁸⁸ Re, and ^(99m) Tc and indirectly with chelating complexes such as diaminetrimethylenepentaacetic acid using ¹¹¹ In, ⁹⁰ Y, ^(99m) Tc, ¹⁸⁶ Re and ¹⁸⁸ Re (47, 48). Antibody-mediated radiotherapy may be carried out using either beta emitting radionuclides such as ¹⁸⁶ Re, ¹⁸⁸ Re, ¹³¹ I, ⁹⁰ Y, ¹⁵³ Sm, ³² P or ¹⁰⁹ Pd, or with alpha particle emitters such as ²¹¹ At or ²¹² Pb, or with Auger electron emitters such as ¹²⁵ I or ¹²³ I (47, 48). Therapy may also be attempted with either drug or toxin based conjugates for example Adriamycin-immunoconjugates (49) and vinblastine-immunoconjugates (50) have been developed. An unexpected finding of the clinical usefulness of immunoscintigraphy has been the reported complete remission of 7 out of 10 FIGO IV ovarian cancer patients who under went repeated imaging with an OC 125 antibody and had anti-idiotypic HAMA (51).

Magnetic resonance imaging can be carried out using gadolinium and other lanthanides or metals such as iron conjugated to antibody based proteins. Several versions of these antibody based products are believed to be undergoing clinical evaluation and commercial development presently.

To improve the specific activity and safety of the immunoconjugate directed towards tumor markers several approaches have been taken ranging from the use of antibody fragments to genetic engineering of recombinant produced humanized antibody constructs to synthetic peptides based on the antibody epitope (52, 53). Methods of antibody engineering including single chain antibodies have been well summarized by Borrebaeck (54). These improvements have improved the tumor target to background ratio and reduced the incidence of human antimouse antibody response.

The present invention provides a method for detecting, locating and treating non-prostatic endocrine tumors involving PSA as the tumor marker and optionally further optionally improves this method by first priming the endocrine tumors to produce PSA.

SUMMARY OF THE INVENTION

According to aspects of the invention, a method for detecting and locating non-prostatic endocrine cancers in vivo by injecting the human subject parenterally with an entity that has been constructed to target PSA, that is either a polyclonal or monoclonal antibody, or fragments thereof, or constructs thereof including but not limited to, single chain antibodies, bifunctional antibodies, molecular recognition units, and peptides or entities that mimic peptides, where the tumor targeting agent is labelled either directly, or indirectly with a chelating agent, with one of ¹³¹ I, ¹²⁵ I, ¹²³ I, ¹¹¹ In, ^(99m) Tc, ⁹⁰ Y, ¹⁸⁶ Re, ¹⁸⁸ Re, ³² P, ¹⁵³ Sm, ⁶⁷ Ga, ²⁰¹ Tl, ⁷⁷ Br or ¹⁸ F and is imaged with a photoscanning device, or where the tumor targeting agent is labelled with either gadolinium, terbium, tin, iron, or isotopes thereof and attached covalently to create a paramagnetic conjugate for the purpose of magnetic resonance imaging.

According to a further aspect of the invention, an in vivo method for imaging endocrine cancer in non-prostatic tissue of a patient comprises:

injecting a patient with biological binding units which bind to PSA produced by non-prostatic tissue of the patient, said PSA-binding units being labelled with imaging agents;

allowing said binding units to incubate in vivo and bind PSA associated with the endocrine cancer; and

detecting presence of said imaging agents of bound units localized to said endocrine cancer.

According to another aspect of the invention, a method for detecting and locating endocrine cancers as described above, wherein the human subject is first given a steroid which induces the cancer cells to express the PSA gene.

Such method further comprises the initial step of injecting a patient with a steroid which induces the cancer cells to produce PSA, said cancer cells having receptors for the injected steroid.

According to another aspect of the invention, a method where entities constructed to target PSA as described above deliver a toxic agent which is a radioisotope that emits Auger electrons, and/or other sub-atomic particles, or toxic compounds including, but not limited to, diphtheria toxin, ricin toxin, adriamycin, chlorambucil, or daunorubicin.

According to another aspect of the invention, a method of passive immunotherapy to endocrine cancer where PSA antibodies, or constructs thereof including, but not limited to, chimeric or human antibodies, or their fragments, single chain antibodies, molecular recognition units, and peptides or entities that mimic peptides are administered parenterally.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is high performance liquid chromatography (HPLC) with a gel filtration column. Each HPLC fraction (0.5 mL) was analyzed with an assay that measures free and a₁ -antichymotrypsin-bound PSA (ACT-PSA) () or an assay that measures only ACT-PSA (♦). The response of the latter assay is in arbitrary fluorescence units since no ACT-PSA standard exists. A. Injection of purified seminal PSA which elutes at fraction 39 corresponding to a molecular weight of 33 kDa. No ACT-PSA is detected. B. Injection of a breast extract from the woman receiving the oral contraceptive Brevicon®. The PSA assay detects two peaks, one at fraction 39 (free PSA, major peak) and one at fraction 30 (100 kDa, minor peak). The latter peak is ACT-PSA as confirmed by the ACT-PSA assay. The identity of the minor peak at fraction 21 (650 kDa) is unknown. This data confirms that over 80% of the breast tissue PSA is in the free, 33 kDa form. The HPLC column was calibrated with molecular weight standards eluting at fraction 21 (660 kDa); 28 (160 kDa); 37 (44 kDa); 42 (17 kDa) and 49 (1.4 kDa).

FIG 2 is a Western blot analysis. Samples were electrophoresed on 8 to 16% gradient polyacrylamide minigels under reducing conditions, electrotransferred to nitrocellulose membranes and probed with a rabbit polyclonal anti-PSA antibody. Detection was achieved by using a horseradish peroxidase-conjugated goat anti-rabbit antibody and chemiluminescence. Lane 1. Molecular weight markers. Lane 2. Purified seminal PSA dissolved in bovine serum albumin. The PSA band appears at 33 koa (just above the 31 kDa marker). Lane 3. Supernatant from a prostatic carcinoma cell line (LNCaP) producing PSA. Lane 4. PSA-positive normal breast extract from the woman receiving Brevicon, containing a band at 33 kDa. Lane 5. Another normal breast extract tested negative for PSA by the immunofluorometric procedure. Lane 6. An amniotic fluid tested for comparison.

FIG. 3 is production of PSA by the breast carcinoma cell line MCF-7. Cells were grown to confluency and then stimulated with varying concentrations of either norethindrone (1) or ethinyl estradiol (2) at the final concentrations indicated, in the absence of fetal calf serum from the culture medium. PSA was measured in the culture supernatant 10 days post No PSA was detected in cell cultures grown identically but either non-stimulated or stimulated with the solvent alone (ethyl alcohol). Norethindrone stimulates PSA production at concentrations as low as 10⁻¹⁰ M.

FIG. 4 is the gamma camera image of a female SCID mouse the left leg of which was injected with norgestrel-stimulated T47-D human breast cancer cells. The image was obtained 21 hours after injection of 10 MBq technetium-99m-labelled B80 anti-PSA monoclonal antibody via the tail vein. The image is an anterior view with the head at the top and the left leg extended to the side.

FIG. 5 is the gamma camera image of a female SCID mouse the left leg of which was injected with non-stimulated T47-D human breast cancer cells. The image was obtained 21 hours after injection of 10 MBq technetium-99m-labelled B80 anti-PSA monoclonal antibody via the tail vein. The image is an anterior view with the head at the top and the left leg extended to the side.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

This invention originated with the surprising discovery that the presence of PSA in human female breast is indicative of breast tumour. PSA was never thought to exist in females because PSA has always been thought to be associated with the male prostate. This discovery and its use in the prognosis of female breast cancer is described in applicant's international application PCT CA94/00267 filed May 13, 1994.

From the discovery of PSA in breast tumors, we have now determined that cancer cells with steroid receptors can be stimulated to produce PSA. It was discovered that normal breast tissue cells and non-PSA producing breast tumor cell lines (our earlier British patent application S.N. 9401491.6 filed Jan. 26, 1994) could be stimulated to produce PSA. Furthermore, non-PSA producing ovarian tumor cells could also be stimulated to produce PSA. Hence, the existence of PSA in a localized region of the body is indicative of a cancer tumor. Procedures to then image the localized concentration of PSA are therefore very useful in cancer diagnosis and prognosis.

Prior to the present invention, agents developed to target PSA were used for diagnostic and therapeutic purposes for prostate cancer only. This invention now provides for the use of radioisotopic or non-isotopic elements containing immunoconjugates directed against PSA in tumours for targeting non-prostatic endocrine cancer in vivo. The procedure is also used for the localization and monitoring of metastases by either nuclear-based gamma camera or magnetic resonance imaging. Another advantage of the invention is the application of reagents directed against PSA labelled with a therapeutically effective radionuclide, drug or toxin for the purpose of therapeutic intervention of breast cancer.

Antibodies or chemical entities created to recognize PSA are used to carry elements to image PSA secreting cancer cells and locate the disease. These antibodies or chemical entities are included in the term biological binding unit, which term is used to refer to patient compatible entities which bind to PSA and comprise antibodies or their derivatives, molecular recognition units and peptides. Antibodies encompass monoclonal and polyclonal antibodies and their derivatives and fragments and include single chain antibodies, bifunctional antibodies and other antibody constructs. Further, these biological binding units may deliver particle emitting radionuclides, drugs or toxins to promote a therapeutic effect. For example, a peptide created to recognize PSA delivering ¹⁸⁸ Re to the tumour site thereby delivers localized radiation to ablate the disease.

By way of a further example, a peptide may be developed that mimics the epitope for anti-PSA and binds to PSA and PSA receptors. This peptide may be produced on a commercially available synthesizer, using FMOC solid phase chemistry. In one application, either tyrosine, lysine, or phenylalanine is included in the peptide to which an N₂ S₂ chelate is complexed as per U.S. Pat. No. 4,897,255. The anti-PSA peptide conjugate is then combined with a radiolabel, for example, either sodium ^(99m) Tc pertechnetate (Na^(99m) TcO₄) or sodium ¹⁸⁸ Re perrhenate(Na¹⁸⁸ ReO₄) and may be used to locate a PSA producing tumor.

The invention also provides the use of anti-PSA antibodies covalently combined with radioactive, cytotoxic or chemotherapeutic molecules and considers using these antibodies in immunoabsorption procedures to separate benign from malignant cells. Further, the concept of passive immunotherapy with antiidotypic antibodies is now possible.

This invention includes a method for detecting and locating non-prostatic endocrine cancers in vivo by injecting a human subject parenterally with an entity that has been constructed to target PSA, that is either a polyclonal or monoclonal antibody, or fragments thereof, or constructs thereof including, but not limited to, single chain antibodies, bifunctional antibodies, molecular recognition units, and peptides or entities that mimic peptides, where the tumour targeting agent is labelled either directly, or indirectly with a chelating agent, with one ¹³¹ I, ¹²⁵ I, ¹²³ I, ¹¹¹ In, ^(99m) Tc, ⁹⁰ Y, ¹⁸⁸ Re, ¹⁵³ Sm, ⁶⁷ Ga, ³² P, ²⁰¹ Tl, ⁷⁷ Br or ¹⁸ F and is imaged with a photoscanning device, or where the tumour targeting agent is labelled with either gadolinium, terbium, tin, iron or isotopes thereof and attached covalently to create a paramagnetic conjugate for the purpose of magnetic resonance imaging. A further application of the radioimaging technique is in the field of radioimmunoguided surgery, whereby a hand-held scintigraphic probe detector enables a surgeon to identify and remove localized metastatic disease (60).

A list of radioisotopes which can be used in the above application is as follows: ²²⁷ Ac, ²¹¹ At, ¹²⁸ Ba, ¹³¹ Ba, ⁷ Be, ²⁰⁴ Bi, ²⁰⁵ Bi, ²⁰⁶ Bi, ⁷⁶ Br, ⁷⁷ Br, ⁸² Br ¹⁰⁹ Cd, ⁴⁷ Ca, ¹¹ C, ¹⁴ C, ³⁶ Cl, ⁴⁸ Cr, ⁵¹ Cr, ⁶² Cu, ⁶⁴ Cu, ⁶⁷ Cu, ¹⁶⁵ Dy, ¹⁵⁵ Eu, ¹⁸ F, ¹⁵³ Gd, ⁶⁶ Ga, ⁶⁷ Ga, ⁶⁸ Ga, ⁷² Ga, ¹⁹⁸ Au, ³ H, ¹⁶⁶ Ho, ¹¹¹ In, ^(113m) In, ^(115m) In, ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸⁹ Ir, ^(191m) Ir, ¹⁹² Ir, ¹⁹⁴ Ir, ⁵² Fe, ⁵⁵ Fe, ⁵⁹ Fe, ¹⁷⁷ Lu, ¹⁵ O, ^(191m-191) Os, ¹⁰⁹ Pd, ³² P, ³³ P, ⁴² K, ²²⁶ Ra, ¹⁸⁶ Re, ¹⁸⁸ Re, ^(82m) Rb, ¹⁵³ Sm, ⁴⁶ Sc, ⁴⁷ Sc, ⁷² Se, ⁷⁵ Se, ¹⁰⁵ Ag, ²² Na, ²⁴ Na, ⁸⁹ Sr, ³⁵ S, ³⁸ S, ¹⁷⁷ Ta, ⁹⁶ Tc, ^(99m) Tc, ²⁰¹ Tl, ²⁰² Tl, ¹¹³ Sn, ^(117m) Sn, ¹²¹ Sn, ¹⁶⁶ Yb, ¹⁶⁹ Yb, ¹⁷⁵ Yb, ⁸⁸ Y, ⁹⁰ Y, ⁶² Zn, ⁶⁵ Zn.

These entities which are constructed to target PSA, as aforementioned, can also deliver a toxic agent for therapeutic purposes against breast cancer, where the toxic agent is a radioisotope that emits Auger electrons, and/or α particles, and/or β particles, and/or neutrons, and/or other sub-atomic particles, or toxic compounds including but not limited to, diphtheria toxin, ricin toxin, adriamycin, chlorambucil, or daunorubicin. Further toxins which can be used are ricin and its derivatives and fragments, Monensin, Verrucarin A, Abrin and its derivatives, Vinca alkaloids, Tricothecenes, and Pseudomonas exotoxin A. Further drugs for use as toxic agents are as follows: Leucovorin, Folinic acid, Methotrexate, Mitomycin C, Neocarzinostatin, Vinblastine, Mitomycin, Melphalan, Mechlorethamine, Fluorouracil, Fluoxuriding, Idarubicin, Doxorubicin, Epirubicin, Cisplatin, Carmustine, Cyclophosphamide, Bleomycin, Vincristine and Cytarabine.

A list of radioisotopes, which can be used for treating endocrine cancers, is as follows: ²²⁷ Ac, ²¹¹ At, ¹³¹ Ba, ⁷⁷ Br, ¹⁰⁹ Cd, ⁵¹ Cr, ⁶⁷ Cu, ¹⁶⁵ Dy, ¹⁵⁵ Eu, ¹⁵³ Gd, ¹⁹⁸ Au, ¹⁶⁶ Ho, ^(113m) In, ^(115m) In, ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸⁹ Ir, ^(191m) Ir, ¹⁹² Ir, ¹⁹⁴ Ir, ⁵² Fe, ⁵⁵ Fe, ⁵⁹ Fe, ¹⁷⁷ Lu, ¹⁰⁹ Pd, ³² P, ²²⁶ Ra, ¹⁸⁶ Re, ¹⁸⁸ Re, ¹⁵³ Sm, ⁴⁶ Sc, ⁴⁷ Sc, ⁷² Se, ⁷⁵ Se, ¹⁰⁵ Ag, ⁸⁹ Sr, ³⁵ S, ¹⁷⁷ Ta, ^(117m) Sn, ¹²¹ Sn, ¹⁶⁶ Yb, ¹⁶⁹ Yb, ⁹⁰ Y, ²¹² Bi, ¹¹⁹ Sb, ¹⁹⁷ Hg, ⁹⁷ Ru, ¹⁰⁰ Pd, ^(101m) Rh, ²¹² Pb.

Since PSA was found to be associated with more benign breast tumors, it was possible that PSA could be expressed by normal breasts either under physiological circumstances or after steroid hormone stimulation, such as described in the aforesaid British patent application. Cytosolic extracts were prepared as previously described (56) from eighteen normal breast tissues removed from nine woman (left and right breast) during breast reduction surgery. PSA immunoreactivity was measured in these extracts using a highly specific and sensitive immunofluorometric technique (57) and by two widely used commercial PSA assays. Breast extracts from eight of the nine women were found to contain <0.03 ng of PSA per mg of total protein and were considered negative for PSA. Surprisingly, two breast extracts from the same woman (left and right breast) had relatively high concentrations of PSA (0.11 and 1.53 ng/mg). None of the eight PSA-negative women was receiving oral contraceptives or other medications. The women with PSA-positive breasts was receiving only one medication, Brevicon®, a highly prescribed oral contraceptive containing 1 mg norethindrone (a progestin) and 0.035 mg ethinyl estradiol per tablet. The PSA-positive and negative results in the breast extracts by the immunofluorometric procedure were verified by using two widely used commercial PSA methods, namely, the IMx® from Abbott Labs, Abbott Park, Chicago, Ill. and the Tandem®-E kit from Hybritech Inc., San Diego, Calif. Additionally, one highly positive extract was serially diluted in female serum from 2- to 32-fold and analyzed by immunofluorometry and the IMx assay. Identical results were obtained.

The highly positive breast extract was also subjected to high performance liquid chromatography (FIG. 1) and fractions were analyzed by two immunofluorometric procedures which measure either total PSA (free PSA plus PSA bound to a₁ -antichymotrypsin) or specifically the PSA-a₁ -antichymotrypsin (ACT) complex (S). Over 80% of the total PSA in normal breast was in the free, 33 kDa form; a small proportion was present as PSA-ACT complex (100 kDa). Another minor species, containing PSA and ACT was also detected (660 kDa), but its identity is unknown. The presence of PSA in the highly positive breast tumor extract was further confirmed by Western blot analysis (FIG. 2). This 33 kDa form of PSA, present in normal breasts stimulated by oral contraceptives, is similar to the PSA form found in breast tumors (1). In male serum, the majority of PSA is present as PSA-ACT complex with a molecular weight of 100 kDa (57).

In order to study the oral contraceptive-induced PSA production further, T-47D and MCF-7 breast carcinoma cell lines were cultured in the absence of any steroid hormones or in the presence of norethindrone or ethinyl estradiol at various concentrations (FIG. 3). No PSA was detected in the tissue culture supernatants in the absence of steroid hormones after 11 days of confluent cultures. Ethinyl estradiol stimulated low levels of PSA production at concentrations ≧10⁻⁸ M. Norethindrone was effective in mediating intense PSA gene expression at concentrations as low as 10⁻¹⁰ M. Other progestins were also effective in mediating PSA gene expression. The identity of PSA in the tissue culture supernatants was further characterized by HPLC and Western blot analysis as shown in FIGS. 1 and 2. Additionally, we were able to amplify by reverse transciption-polymerase chain reaction (RT-PCR), prostate specific antigen mRNA from the stimulated but not the non-stimulated cells and verify its identify by Southern hybridization and sequencing of the PCR product (58). The same procedures confirmed the presence of PSA in breast tumors positive for the protein.

We recently demonstrated that among 99 ovarian tumor extracts tested only three were positive for PSA and the concentration of the PSA was 0.048, 0.034 and 0.0076 ng/mg. Subsequently, a patient with ovarian cancer who also underwent liver transplantation and was receiving oral prednisone tablets during the period of ovarian tumor removal was tested for the presence of PSA in the ovarian tumor. Remarkably, this ovariant tumor contained 15 ng of PSA/mg protein, which is a very high amount. Combined with our previous results on breast tumor cell line tissue culture systems (British Patent Application S.N. 9401491.7) in which we found that glucocorticosteroids can stimulate PSA production, it is apparent that the ovarian tumor, generally unable to produce PSA, can be stimulated by steroids like prednisone, to produce very high levels of PSA in the tumor. The tumors, which may be imaged and treated in this invention, have steroid receptors which are stimulated in the presence of steroids to express the PSA gene and thus produce PSA. The PSA-binding units can then bind to cancer cells producing PSA which allows these cancers to be treated and localized.

Following this demonstration of stimulation of PSA gene expression in breast carcinoma cell lines and ovarian tumors, it is a further feature of the invention that stimulating cancers to express PSA would either enable detection of previously undetectable tumors or would improve radioimaging of previously detectable tumors. Indeed as shown with the normal breast tissue of the patient receiving the oral contraceptive, steroid receptor positive tissues could be induced to produce PSA which would enable these tissues to be radioimaged. Further, the production of PSA by the non-PSA producing normal breast and ovarian tumor tissues after stimulation will enable radioimaging of non-PSA producing endocrine tumors after stimulation. While normal endocrine tissue can be stimulated to produce PSA, it will not be produced at the high levels produced by stimulated endocrine cancer cells. Thus, priming a patient with a steroid which induces PSA gene expression to, in turn, produce PSA at the tumor site, provides increased binding of the entity targeting PSA to endocrine tumors and improves the radioimaging or therapeutic delivery of toxic agents.

Gamma camera images of SCID mice with norgestrel-stimulated (FIG. 4) and non-stimulated (FIG. 5) T47-D human breast cancer cells injected into the muscle of the left leg were taken. Images were obtained 21 hours after injection of 10 MBq technetium-99m-labelled B80 anti-PSA monclonal antibody via the tail vein. Images are anterior views, heat at top, with left leg, which contains the tumor cells, extended to the side and immobilized. An upper threshold of 5% was used to mask the residual radioactivity in the liver and the abdomen. (That is, the top 5% of counts were subtracted from all images to allow the leg tumor to be viewed.) The leg containing the tumor is seen in the lower right portion of each image. Quantification of this pair of digital images showed that the stimulated tumor contained at least 15% more radioactivity than the control tumor.

Tissue counting results were obtained for blood, normal muscle and either the stimulated or control T47-D tumor and are provided in Table 1. All values are percent injected dose per gram tissue, expressed as a mean ±standard deviation for 4 animals or individual values for 2 animals. Table 1 demonstrates that the T47-D tumor cells had at least double the counts of the control. Furthermore, the stimulated tumor cells have increased counts over that of the non-stimulated tumor cells. Thus radioimaging was improved by stimulation to increase PSA production.

                  TABLE 1                                                          ______________________________________                                         Blood            0.96 ± 0.14                                                Normal muscle    0.129 ± 0.029                                              T47-D control tumor                                                                             0.228-0.231                                                   (range 2 animals)                                                              T47D- stimulated tumor                                                                          0.266-0.405                                                   (range 2 animals)                                                              ______________________________________                                    

It was proven that the norgestrel-stimulated T-47D cells in the above experiment were producing PSA from the detection of PSA in the cell culture supernatant. Furthermore, T-47D and MCF-7 tumor cell lines were injected into SCID mice to develop tumors, which mice were then injected with estrogen and/or norgestrel. The results (Table 3) demonstrated that human PSA was found in the serum of mice injected with norgestrel, however, estrogen blocked the effect of the norgestrel. Estrogen was previously shown to block the effect of progestin on PSA production in the tumor cell lines tested in our earlier British patent application S.N. 9401491.7.

The breast tumor cell line tissue culture system (described in British patent application S.N. 9401491.7) suggested that any steroid having either glucocorticoid, mineralocorticoid, antiestrogen, progestin or androgen activity can regulate the PSA gene; however, estrogens cannot mediate such action. These results combined with the results from steroid stimulation of PSA expression in normal breast cells and non-PSA producing breast tumor cell lines and ovarian tumor, indicate that to stimulate tumors for radioimaging applications, any natural or synthetic steroid falling under the above categories of activity would be suitable. A list of steroids to induce expression of the PSA gene is found in Table 2.

A list of non-prostatic endocrine tumors which can be stimulated to express the PSA gene either initially or to increase PSA expression is as follows: breast tumors, ovarian tumors, lung carcinomas, meningiomas, endometrial carcinomas, colon carcinomas, salivary gland tumors, cervical carcinomas, uterine carcinomas, adrenal tumors, renal carcinomas and melanomas. The steroids, which stimulate the production of PSA, may be naturally present in the body or may be introduced artificially by injection.

An alternative aspect of this invention, as it relates to in vivo antibody binding, is a method of passive immunotherapy to non-prostatic endocrine cancer where PSA antibodies, or constructs thereof including, but not limited to, chimeric or human antibodies, or their fragments, single chain antibodies, molecular recognition units, and peptides or entities that mimic peptides are administered parenterally. Antiidiotypic antibodies may then be produced in reaction to this administration.

Antiidiotypic antibodies are antibodies directed against the idiotype (the antigen combining region or variable region) of another antibody, in this case anti-PSA antibody. These antiidiotypic antibodies in turn stimulate the production of antibodies against the idiotype of the antiidiotypic antibodies. Thus this sub-population of antiidiotypic antibodies bind the initial antigen, PSA. For example, human anti-PSA antibodies can be administered to patients that have PSA secreting breast tumors. A dose escalating regime can be followed, over a period of months, if there is an absence of any untoward response. Patient sera can be examined via in vitro methods for the development of antiidiotypic antibodies. The development of anti-idiotypic antibodies results in enhancement or beneficial modification of the patient's immune response and thereby elicits an anti-tumor response (51).

EXAMPLE 1

Breast tumors were snap-frozen in liquid nitrogen. Frozen sections (5 μm thick) were placed on clean glass slides and processed for immunohistochemistry. Slides to be stained for PSA were fixed immediately in 10% buffered formalin for 10 min. and then placed in phosphate-buffered saline. Staining was performed with a polyclonal anti-PSA antibody from Medix Biotech, Foster City, Calif. and further completed by use of the avidin-biotin technique (ABC) using a kit from Vector Laboratories, Burlingame, Calif. Immunoperoxidase staining was, according to the manufacturer's recommendation (58). Tumors positive or negative for PSA by the immunofluorometric procedure were used. The slides were examined under a light microscope. Tumors, negative for PSA showed no staining while tumors positive for PSA exhibited strong brown cytoplasmic staining with the immunoperoxidase technique. This experiment demonstrated that PSA in breast tumors can be localized by antibodies against PSA.

EXAMPLE 2

Cell Preparation

The following cell line was obtained from the ATCC: T47-D, an estrogen dependent cell line derived from breast cancer. The cell line was checked for freedom from microbial contaminants and all operations were carried out aseptically. T47-D cells were cultured in α Medium with 10% fetal calf serum in sterile culture flasks. Cells were then incubated at 37° C. and growth was checked visually using a microscope. Two sets of T47-D cells were prepared, one set stimulated with norgestrel, a progestin, and the other set non-stimulated. The stimulated set were stimulated with a solution of norgestrel (concentration 1 millimole in ethanol), at the concentration of 1 micromolar in fresh media for a period of 48 h. PSA was detected in the supernatant of the norgestrel-stimulated cells' supernatant after 72 h and the concentration of PSA increased from 0.013 μg/L to 0.037 μg/L at 96 h.

Cells were scraped into a centrifuge tube and spun down in a laboratory centrifuge at approximately 240 G for six minutes. Cells were re-suspended using the sterile culture medium and re-spun, the procedure was repeated a total of three times. For a 0.2 ml pellet of cells, 0.1 ml was injected subcutaneously into the left leg, slightly below the knee, in female SCID mice. The amount of cells injected into the SCID mice were estimated to be between 2.5×10⁶ and 1×10⁷. The experiment was repeated once using trypsin to avoid clumping of the harvested cells, in which case the stimulation with norgestrel was increased to 2 micromolar in fresh culture medium to overcome any effect due to trypsin. The results of imaging shown in FIGS. 4 and 5 were obtained using cells harvested by scraping, whereas the results of Table 1 were obtained using cells harvested with trypsin.

Preparation of the Monoclonal Antibody

The anti-PSA monoclonal antibody, B 80, was obtained from Biomira Inc., Edmonton, Alberta. The antibody was prepared for radioimaging using the method described by Joiris et al. (55) with 2 iminothiolane. Three solutions are first prepared: 5 milligram/ml solution of B 80 in physiological saline; 10 milligram/ml solution of 2 iminothiolane; and a phosphate buffer solution containing 0.25M of phosphate adjusted to pH 7.4. 10 micorliters of each of these solutions was combined and allowed to stand for 30 minutes at about 25° C. This solution was then combined with 100 microliters of ^(99m) TcO₄ ⁻ (approximately 10 mCi/ml or 370.4 MBq/mL) and 100 microliters of a dilute stannous glucoheptonate solution containing 5 micrograms of stannous chloride. This solution was allowed to incubate for 10 minutes at 25° C. The solution was then purified using a Sephadex G-50 column which has been preblocked with a 1% solution of human serum albumen in 0.9% NaCl. The column was eluted using 0.9% NaCl. The hottest fraction, as determined by measurements using a Capintec Dose Calibrator was kept and diluted to 50 Mbq/ml using 0.9% NaCl. 0.2 ml, a solution equal to 10 MBq was the injected dose.

Imaging Studies

0.2 ml of ^(99m) Tc radiolabelled B 80 antibody was injected intravenously into the tail vein of the SCID mice prepared as described above. Imaging studies were carried out using a Siemans Orbiter, model 7400 gamma camera with a low energy all purpose collimator using a magnification factor of 2.5. Data was collected using a Picker PCS I-II nuclear medicine computer. Counts were obtained using a 128×128 matrix. Imaging studies were carried out with the mouse conscious in a lucite restraining device with the leg containing the tumor taped to the base plate. The restraining device was placed directly on the collimator. Initial images were carried out between 1 and 7 hours post injection using a five minute data acquisition and repeated at 20 hours using a 15 minute acquisition. Quantification was performed by drawing a region of interest around the tumor. After correction for background counts, comparisons of the norgestrel-stimulated and control tumors were made using counts per pixel values.

Following the final imaging session, the mice were sacrificed by cervical dislocation and samples of blood, leg muscle containing tumor cells, and control muscle from the opposite leg were obtained, weighed and assayed in a gamma well counter (Capintec). From the knowledge of the dose administered and counting efficiency, the percent dose per gram was calculated.

EXAMPLE 3

T-47D tumor cells or MCF-7 tumor cells were implanted into SCID mice and left to develop as tumors over several weeks. Mice were injected subcutaneously with estrogen and/or norgestrel in ethanol (100 μL of a 10⁻³ M solution per mouse) and serum was collected after 48 hours. The results from measuring PSA within the serum by the fluorescence method (57) are found in Table

                  TABLE 3                                                          ______________________________________                                                                             Serum PSA                                                                      Assay,                                     Experiment                                                                              Tumor Type Estrogen Norgestrel                                                                            Fluorescence                               ______________________________________                                         1        T-47D      -        +      2818                                       2        T-47D      +        +      2661                                       3        MCF-7      +        +      1912                                       4        MCF-7      -        +      6893                                       5        MCF-7      +        -      2067                                       6        MCF-7      -        +      2642                                       ______________________________________                                    

                  TABLE 2                                                          ______________________________________                                         LIST OF STEROIDS                                                               3β-Acetoxy-9(11),16-allopregnadien-20-one                                 21-Acetoxyallopregnan-3,20-dione                                               3β-Acetoxy-5-androsten-17-one                                             3β-Acetoxybisnorallocholanic acid                                         3β-Acetoxybisnor-5-cholenic acid                                          21-Acetoxy-3α,17-dihydroxy-5β-pregnan-11,20-dione                   3α-Acetoxy-5β-etianic acid                                          3β-Acetoxy-5β-etianic acid                                           3β-Acetoxy-5-etienic acid                                                 3β-Acetoxyetiocholenic acid                                               21-Acetoxy-17-hydroxyallopregnan-3,11,20-trione                                3β-Acetoxy-5,16-pregnadien-20-one                                         12α-Acetoxypregnan-3,20-dione                                            21-Acetoxypregnanedione                                                        Acetoxypregnanolone                                                            17-Acetoxypregnenolone                                                         21-Acetoxypregnenolone                                                         3β-Acetoxy-16,(5β)-pregnen-20-one                                    11α-Acetoxyprogesterone                                                  17-Acetoxyprogesterone                                                         17-Acetoxyprogesterone 3-ethyleneketal                                         21-Acetoxyprogesterone                                                         Δ1-Adrenosterone                                                         Adrenosterone                                                                  Aetiocholane                                                                   Aldosterone                                                                    Aldosterone 21-acetate                                                         Aldosterone 3-CMO                                                              Aldosterone 3-CMO:BSA                                                          Aldosterone 18,21-diacetate                                                    Aldosterone diacetate 3-CMO                                                    Aldosterone diacetate 3-CMO:BSA                                                Aldosterone 21-hemisuccinate                                                   Aldosterone 21-hemisuccinate:BSA                                               Allocholesterol                                                                Allodihydrocorticosterone                                                      Allodihydrocortisol                                                            Allodihydrocortisone                                                           Allodihydrocortisone acetate                                                   Allodihydro substance "S"                                                      Allodihydrotestosterone                                                        Allopregnanediol                                                               Allopregnanolone                                                               Allopregnanolone acetate                                                       Allotetrahydro compound "A"                                                    Allotetrahydro compound "B"                                                    Allotetrahydro compound "E"                                                    Allotetrahydro compound "F"                                                    Allotetrahydrocorticosterone                                                   Allotetrahydrocortisol                                                         Allotetrahydrocortisone                                                        Allotetrahydro-11-dehydrocorticosterone                                        Allotetrahydrodesoxycorticosterone                                             Allotetrahydrodesoxycorticosterone 21-acetate                                  Allotetrahydro-11-desoxycortisol                                               Allotetrahydro DOC                                                             Allotetrahydro DOC 21-acetate                                                  Allotetrahydrohydrocortisone                                                   Allotetrahydro substance "Q"                                                   Allotetrahydro substance "S"                                                   Allo TH "A"                                                                    Allo TH "B"                                                                    Allo TH "E"                                                                    Allo TH "E" diacetate                                                          Allo TH "F"                                                                    Allo TH "F" diacetate                                                          Allo Th "S"                                                                    Androstadienedione                                                             Androstadienedione 3-ethylenol ether                                           Androstanedione                                                                Androstanolone                                                                 Androstatriendione                                                             Androstenediol                                                                 Androstenedione                                                                Androstenolone                                                                 4-Androsten-17β-ol-one enol diacetate                                     Androsterone                                                                   Androsterone acetate                                                           Androsterone benzoate                                                          Androsterone chloroformate                                                     Androsterone-CMO                                                               Androsterone glucuronide                                                       Androsterone hemisuccinate                                                     Androsterone propionate                                                        Androsterone sodium sulfate                                                    Androsterone tosylate                                                          Anhydroxyprogesterone                                                          Apocholic acid                                                                 Beclomethasone                                                                 Beclomethasone 21-acetate                                                      Beclomethasone 17,21-dipropionate                                              Beclomethasone 21-hemisuccinate                                                Beclomethasone 21-propionate                                                   Betamethasone                                                                  Betamethasone 21-acetate                                                       Betamethasone-17,21-dipropionate                                               Betamethasone 21-disodium phosphate                                            Betamethasone 21-hemisuccinate                                                 Betamethasone 17-valerate                                                      3,4-Bis(4-hydroxyphenyl)-hexane                                                3,4-Bis-(4-hydroxyphenyl)-3-hexene                                             Bolderone                                                                      Bolderone sulfate, sodium salt                                                 2α-Bromo-5α-cholestan-3-one                                        4-Bromoequilenin                                                               4-Bromoequilin                                                                 2-Bromoestradiol                                                               4-Bromoestradiol                                                               16α-Bromoestradiol                                                       16α-Bromo-17α-estradiol                                            16α-Bromoestrone                                                         16β-Bromoestrone                                                          2-Bromoethynylestradiol                                                        4-Bromoethynylestradiol                                                        17-Bromopregnenolone                                                           17-Caproxyprogesterone                                                         Chenodeoxycholic acid                                                          Chenodeoxycholic acid 3-Hemisuccinate                                          Chenodeoxycholic acid methyl ester                                             Chenodeoxycholic acid sodium salt                                              3β-Chloro-5α-cholestane                                             3β-Chloro-5-cholestene                                                    21-Chloro-17-hydroxyprogesterone                                               Cholaic acid                                                                   α-Cholestanol                                                            β-Cholestanol                                                             Cholestanol                                                                    Cholestanol acetate                                                            Cholestanol benzoate                                                           Cholestanol hemisuccinate                                                      Cholestanol propionate                                                         Cholestanol tosylate                                                           5α-Cholestan-3β-ol chloride                                         Cholestanone                                                                   5α-Cholestan-3-one enol acetate                                          5α-Cholestan-3β-yl chloride                                         Cholestanyl chloride                                                           Cholestene                                                                     Cholestenol                                                                    5-Cholesten-3β-ol chloride                                                Cholestenone                                                                   4-Cholesten-3-one enol acetate                                                 Cholesterilene                                                                 Cholesterin                                                                    Cholesterol                                                                    Cholesteryl acetate                                                            Cholesteryl acetoacetate                                                       Cholesteryl benzoate                                                           Cholesteryl n-butyrate                                                         Cholesteryl caprylate                                                          Cholesteryl chloride                                                           Cholesteryl chloroacetate                                                      Cholesteryl chloroformate                                                      Cholesteryl cinnamate                                                          Cholesteryl n-decylate                                                         Cholesteryl ethylether                                                         Cholesteryl formate                                                            Cholesteryl glucuronide, sodium salt                                           Cholesteryl hemisuccinate                                                      Cholesteryl heptanoate                                                         Cholesteryl hexadecanoate                                                      Cholesteryl hydrocinnamate                                                     Cholesteryl hydrogen phthalate                                                 Cholesteryl iso-butyrate                                                       Cholesteryl laurate                                                            Cholesteryl methylcarbonate                                                    Cholesteryl methyl ether                                                       Cholesteryl myristate                                                          Cholesteryl nonanoate                                                          Cholesteryl octanoate                                                          Cholesteryl pelargonate                                                        Cholesteryl β-phenylpropionate                                            Cholesteryl n-propionate                                                       Cholesteryl pyridinium sulfate                                                 Cholesteryl sodium sulfate                                                     Cholesteryl stearate                                                           Cholesteryl tosylate                                                           Cholesteryl valerate                                                           Cholic acid                                                                    Cholic acid methyl ester                                                       Cholic acid sodium salt                                                        Cinchol                                                                        Cistestosterone                                                                Coprostane                                                                     Coprostanol                                                                    Coprostanol acetate                                                            Coprostanol benzoate                                                           Coprostan-3-one                                                                Coprostenol                                                                    Coprosterol                                                                    Cortexolone                                                                    Cortexone                                                                      Corticosterone                                                                 Corticosterone acetate                                                         Corticosterone 21-acetate, 3-CMO                                               Corticosterone 21-acetate, 3-CMO:BSA                                           Corticosterone 3-CMO                                                           Corticosterone 3-CMO:BSA                                                       Corticosterone diethyleneketal                                                 Corticosterone hemisuccinate                                                   Corticosterone hemisuccinate:BSA                                               Cortisol                                                                       Cortisol acetate                                                               Cortisol 21-acetate, 3-CMO                                                     Cortisol 21-acetate, 3-CMO:BSA                                                 Cortisol 3-CMO                                                                 Cortisol 3-CMO:BSA                                                             Cortisol glucuronide                                                           Cortisol glucuronide, sodium salt                                              Cortisol hemisuccinate                                                         Cortisol 17-valerate                                                           Δ1-Cortisone                                                             Cortisone                                                                      Cortisone acetate                                                              Cortisone 21-acetate, 3-CMO                                                    Cortisone 21-acetate, 3-ethyleneketal                                          Cortisone diethyleneketal                                                      Cortisone 21-hemisuccinate                                                     Cortisone 21-sodium sulfate                                                    Cortol                                                                         β-Cortol                                                                  Cortolone                                                                      β-Cortolone                                                               DES                                                                            DHEA                                                                           DHEA acetate                                                                   DOCA                                                                           DOCA 3-ethyleneketal                                                           DOC 21-aldehyde hemiacetal                                                     DOC 3-CMO                                                                      DOC 3-CMO:BSA                                                                  DOC glucuronide                                                                DOC hemisuccinate                                                              DOC propionate                                                                 DPA                                                                            16-Dehydroallopregnanolone                                                     16-Dehydroallopregnanolone acetate                                             24-Dehydrocholesterol                                                          Dehydrocholic acid                                                             1-Dehydrocortisone                                                             11-Dehydrocorticosterone                                                       11-Dehydrocorticosterone acetate                                               11-Dehydrocorticosterone hemisuccinate                                         Dehydrocortisol                                                                Dehydroepiandrosterone                                                         Dehydroepiandrosterone acetate                                                 Dehydroepiandrosterone glucuronide                                             Dehydroepiandrosterone potassium sulfate                                       Dehydroepiandrosterone propionate                                              Dehydroepiandrosterone sodium sulfate                                          Dehydroepiandrosterone tosylate                                                6-Dehydro-17α-estradiol                                                  6-Dehydroestradiol                                                             6-Dehydroestradiol diacetate                                                   7-Dehydro-17α-estradiol                                                  7-Dehydro-17β-estradiol                                                   7-Dehydro-17β-estradiol diacetate                                         16-Dehydroestradiol diacetate                                                  6-Dehydroestrone                                                               6-Dehydroestrone acetate                                                       6-Dehydroestrone benzoate                                                      6-Dehydroestrone benzyl ether                                                  6-Dehydroestrone methyl ether                                                  8-Dehydroestrone                                                               Dehydroisoandrosterone                                                         Dehydroisoandrosterone acetate                                                 1-Dehydromethyltestosterone                                                    16-Dehydro-5α-pregnan-3β-ol 11,20-dione                             16-Dehydro-5α-pregnan-3β-ol 11,20-dione acetate                     16-Dehydropregnanolone                                                         16-Dehydropregnanolone acetate                                                 16-Dehydro-5α-pregnan-3β-ol 20-one                                  16-Dehydro-5α-pregnan-3β-ol 20-one acetate                          16-Dehydro-5β-pregnan-3β-ol 20-one                                   16-Dehydro-5β-pregnan-3β-ol 20-one acetate                           16-Dehydropregnenolone                                                         16-Dehydropregnenolone acetate                                                 16-Dehydropregnenolone acetate, oxime                                          16-Dehydropregnenolone oxime                                                   16-Dehydroprogesterone                                                         1-Dehydrotestosterone                                                          1-Dehydrotesrosterone acetate                                                  1-Dehydrotestosterone benzoate                                                 1-Dehydrotestosterone 3-CMO                                                    1-Dehydrotestosterone hemisuccinate                                            1-Dehydrotestosterone propionate                                               1-Dehydrotestosterone sodium sulfate                                           6-Dehydrotestosterone                                                          6-Dehydrotestosterone acetate                                                  6-Dehydrotestosterone benzoate                                                 6-Dehydrotestosterone 3-CMO                                                    6-Dehydrotestosterone hemisuccinate                                            6-Dehydrotestosterone propionate                                               Deoxycholic acid                                                               Deoxycholic acid diacetate                                                     Deoxycholic acid sodium salt                                                   Desmosterol                                                                    Desmosterol acetate                                                            Desonide                                                                       Desoxycorticosterone                                                           Desoxycorticosterone acetate                                                   Desoxycorticosterone acetate 3-CMO                                             Desoxycorticosterone acetate 3-CMO:BSA                                         Desoxycorticosterone acetate, 3-ethyleneketal                                  Desoxycorticosterone 21-aldehyde hemiacetal                                    Desoxycorticosterone 3-CMO                                                     Desoxycorticosterone 3-CMO:BSA                                                 Desoxycorticosterone glucuronide                                               Desoxycorticosterone hemisuccinate                                             Desoxycorticosterone propionate                                                11-Desoxycortisol                                                              21-Desoxycortisol                                                              21-Desoxycortisone                                                             17-Desoxycortol                                                                17-Desoxy-β-cortol                                                        17-Desoxy-β-cortolone                                                     11-Desoxy-17-hydroxycorticosterone                                             Desoxymethasone                                                                Dexamethasone                                                                  Dexamethasone acetate                                                          Dexamethasone hemisuccinate                                                    Dexamethasone hemisuccinate:BSA                                                Dexamethasone 21-mesylate                                                      Dexamethasone phosphate disodium salt                                          Dianabol                                                                       2,4-Dibromoestradiol                                                           Dichlorisone                                                                   3α,12α-Diformyloxydesoxycholic acid                                Dihydroandrosterone                                                            Dihydrocholesterol                                                             Dihydrocholesterol acetate                                                     Dihydrocholesterol benzoate                                                    Dihydrocholesterol hemisuccinate                                               Dihydrocholesterol methyl ether                                                Dihydrocholesterol propionate                                                  Dihydrocholesterol tosylate                                                    5α-Dihydrocortexone                                                      5α-Dihydrocorticosterone                                                 5α-Dihydrocortisol                                                       5α-Dihydrocortisone acetate                                              20β-Dihydrocorticosterone                                                 20β-Dihydrocortisol                                                       Dihydrocortisone                                                               5α-Dihydrocortisone                                                      5β-Dihydrocortisone                                                       Dihydrocortisone acetate                                                       5α-Dihydro-11-dehydrocorticosterone                                      Dihydro-11-desoxycortisol                                                      17β-Dihydroequilenin                                                      17β-Dihydroequilenin diacetate                                            17α-Dihydroequilenin                                                     17β-Dihydroequilenin                                                      17β-Dihydroequilenin diacetate                                            20β-Dihydro Kendall's compound "B"                                        5α-Dihydro substance "Q"                                                 5α-Dihydro substance "S"                                                 5α-Dihydrotestosterone                                                   Dihydrotestosterone                                                            Dihydrotestosterone acetate                                                    Dihydrotestosterone benzoate                                                   Dihydrotestosterone 3-CMO                                                      Dihydrotestosterone 3-CMO:BSA                                                  Dihydrotestosterone chloroformate                                              Dihydrotestosterone cyclopentylpropionate                                      Dihydrotestosterone enanthate                                                  Dihydrotestosterone glucuronide                                                Dihydrotestosterone hemisuccinate                                              Dihydrotestosterone hemisuccinate:BSA                                          Dihydrotestosterone hexahydrobenzoate                                          Dihydrotestosterone propionate                                                 Dihydrotestosterone tosylate                                                   5β-Dihydrotestosterone                                                    5β-Dihydrotestosterone acetate                                            5β-Dihydrotestosterone 3-CMO                                              5β-Dihydrotestosterone hemisuccinate                                      5β-Dihydrotestosterone propionate                                         3α,12α-Dihydroxycholanic acid                                      5β-Dihydroxycorticosterone                                                3α,7α-Dihydroxy-12-ketocholanic acid                               3α,12α-Dihydroxynorcholanate                                       3β,17-Dihydroxy-5-pregnen-3-one                                           Dihydro Reichstein's substance "S"                                             3,6-Diketocholanic acid                                                        3,7-Diketocholanic acid                                                        3,12-Diketocholanic acid                                                       7,12-Diketolithocholic acid                                                    7,12-Diketolithocholic acid sodium salt                                        6,16-Dimethyl-16-dehydroprogesterone                                           2,4-Dinitroestradiol                                                           Diosgenin                                                                      Diosgenin acetate                                                              Diosgenin benzoate                                                             E 1                                                                            E 2                                                                            E 3                                                                            E 4                                                                            α-Ecdysone                                                               Electrocortin                                                                  Epi-allocholesterol                                                            Epi-allotetrahydro "B"                                                         Epiandrosterone                                                                Epiandrosterone acetate                                                        Epiandrosterone benzoate                                                       Epiandrosterone chloroformate                                                  Epiandrosterone 17-CMO                                                         Epiandrosterone glucuronide                                                    Epiandrosterone hemisuccinate                                                  Epiandrosterone potassium sulfate                                              Epiandrosterone propionate                                                     Epiandrosterone sodium sulfate                                                 Epiandrosterone tosylate                                                       Epicholestanol                                                                 Epicholesterol                                                                 Epi compound "F"                                                               Epicoprostanol                                                                 Epicoprostanol acetate                                                         Epicoprostanol benzoate                                                        Epicoprostanol hemisuccinate                                                   Epicoprostanol propionate                                                      Epicoprosterol                                                                 Epidihydrocholesterol                                                          16-Epiestriol                                                                  16-Epiestriol 3-methylether                                                    16-Epiestriol triacetate                                                       17-Epiestriol                                                                  17-Epiestriol triacetate                                                       16,17-Epiestriol                                                               16,17-Epiestriol triacetate                                                    Epitestosterone                                                                Epitestosterone acetate                                                        Epitestosterone benzoate                                                       Epitestosterone hemisuccinate                                                  11-Epi Tetrahydro compound "B"                                                 11-Epi-Tetrahydro compound "F"                                                 11-Epi-Terrahydrocorticosterone                                                11-Epi-Tetrahydrocortisol                                                      11-Epi-Tetrahydrohydrocortisone                                                11-Epi-TH "B"                                                                  11-Epi-TH "F"                                                                  Epoxypregnanolone                                                              Epoxypregnenolone                                                              Epoxypregnenolone acetate                                                      16α,17-epoxyprogesterone                                                 d-Equilenin                                                                    d-Equilenin acetate                                                            d-Equilenin benzoate                                                           d-Equilenin benzyl ether                                                       d-Equilenin etheyl ether                                                       d-Equilenin methyl ether                                                       Equilin                                                                        Equilin acetate                                                                Equilin benzoate                                                               Equilin methylether                                                            4,22-Ergostadien-3-one                                                         Ergosterol                                                                     Esmilagenin                                                                    Estetrol                                                                       17α-Estradiol                                                            17α-Estradiol 3-acetate                                                  17α-Estradiol 17-acetate                                                 17α-Estradiol diacetate                                                  17β-Estradiol                                                             17β-Estradiol 3-acetate                                                   17β-Estradiol 17-acetate                                                  17β-Estradiol 17-acetate, 3-benzoate                                      17β-Estradiol 17-acetate, 3-methyl ether                                  17β-Estradiol 3-benzoate                                                  17β-Estradiol 3-benzoate, 17-valerate                                     17β-Estradiol 3-benzyl ether                                              17β-Estradiol 17-cyclopentylpropionate                                    17β-Estradiol diacetate                                                   17β-Escradiol dibenzoate                                                  17β-Estradiol dicyclopentylpropionate                                     17β-Estradiol diglucuronide                                               17β-Estradiol dihemisuccinate                                             17β-Estradiol dipalmitate                                                 17β-Estradiol diphosphate, disodium salt                                  17β-Estradiol dipropionate                                                17β-Estradiol disodium sulfate                                            17β-Estradiol 17-enanthate                                                17β-Estradiol 3-glucuronide                                               17β-Estradiol 17-glucuronide                                              17β-Estradiol 3-hemisuccinate                                             17β-Estradiol 17-hemisuccinate                                            17β-Estradiol 17-hemisuccinate:BSA                                        17β-Estradiol 17-hexahydrobenzoate                                        17β-Estradiol 3-methyl ether                                              17β-Estradiol 17-phenylpropionate                                         17β-Estradiol 3-phosphate, disodium salt                                  17β-Estradiol 17-phosphate, disodium salt                                 17β-Estradiol 3-sodium sulfate                                            17β-Estradiol 17-sodium sulfate                                           17β-Estradiol 17-stearate                                                 17β-Estradiol 17-valerate                                                 Estriol                                                                        Estriol 3-acetate                                                              Estriol 16-acetate                                                             Estriol 16,17-diacetate                                                        Estriol 3,17-disodium sulfate                                                  Estriol 16,17-disodium sulfate                                                 Estriol 3-glucuronide, sodium salt                                             Estriol 3-hemisuccinate                                                        Estriol 16-hemisuccinate                                                       Estriol 3-methyl ether                                                         Estriol 3-phosphate, disodium salt                                             Estriol 3-sodium sulfate                                                       Estriol 17-sodium sulfate                                                      Estriol triacetate                                                             Estriol tripropionate                                                          Estrone                                                                        Estrone acetate                                                                Estrone benzoate                                                               Estrone benzyl ether                                                           Estrone 17-enol acetate, 3-methyl ether                                        Estrone enol diacetate                                                         Estrone ethyl ether                                                            Estrone glucuronide, sodium salt                                               Estrone hemisuccinate                                                          Estrone methoxime                                                              Estrone methyl ether                                                           Estrone phosphate, disodium salt                                               Estrone propionate                                                             Estrone trimethylacetate                                                       Ethisterone                                                                    24b-Ethylcholesterol                                                           17α-Ethyl-19-nor-testosterone                                            Ethynodiol                                                                     Ethynylandrostanolone                                                          Ethynylandrostendiol                                                           Ethynylandrostenolone                                                          17α-Ethynyldihydrotestosterone                                           Ethynylestradiol                                                               17α-Ethynylestadiol 3-acetate                                            Ethynylestradiol 3-methyl ether                                                17α-Ethynyl-19-nor-testosterone                                          Ethynyltestosterone                                                            Etiadienic acid                                                                Etiadienic acid 3-acetate                                                      Etiadienic acid methyl ester                                                   Etianic acid                                                                   Etienic acid                                                                   Etienic acid acetate                                                           Etienic acid methyl ester                                                      Etiocholane                                                                    Etiocholan-3α,17α-diol                                             Etiocholan-3α,17β-diol                                              Etiocholan-3α,17β-diol diacetate                                    Etiocholan-3β,17α-diol                                              Etiocholan-3β,17β-diol                                               Etiocholan-3β,17β-diol diacetate                                     Etiocholan-3α,6α-diol-17-one                                       Etiocholan-3β,11β-diol-17-one                                        Etiocholan-3,17-dione                                                          Etiocholan-3α-ol                                                         Etiocholan-3α-ol-11,17-dione                                             Etiocholanolone                                                                Etiocholanolone acetate                                                        Etiocholanolone benzoate                                                       Etiocholanolone 17-CMO                                                         Etiocholanolone glucuronide                                                    Etiocholanolone hemisuccinate                                                  Etiocholanolone potassium sulfate                                              Etiocholanolone propionate                                                     Etiocholanolone sodium sulfate                                                 Etiocholan-3β-ol-17-one                                                   Etiocholan-17β-ol-3-one                                                   Etiocholan-3β-ol-17-one acetate                                           Etiocholan-3β-ol-17-one benzoate                                          Etiocholan-3β-ol-17-one hemisuccinate                                     Etiocholan-3β-ol-17-one propionate                                        Etiocholenic acid acetate                                                      Flucinonide                                                                    Flumethasone                                                                   Fluocinolone acetonide                                                         2-Fluoroestradiol                                                              Fluorometholone                                                                Fluoxymesterone                                                                Flurandrenolide                                                                Flurocortisone                                                                 Fucosterol                                                                     Glycochenodeoxycholic acid                                                     Glycochenodeoxycholic acid sodium salt                                         Glycocholanic acid                                                             Glycocholic acid                                                               Glycocholic acid potassium salt                                                Glycocholic acid sodium salt                                                   Glycodehydrocholic acid                                                        Glycodehydrocholic acid sodium salt                                            Glycodeoxycholic acid                                                          Glycodeoxycholic acid sodium salt                                              Glycohyodeoxycholic acid                                                       Glycohyodeoxycholic acid sodium salt                                           Glycolithocholic acid                                                          Glycolithocholic acid sodium salt                                              Hecogenin                                                                      Hecogenin acetate                                                              Hetero-1-methylestradiol diacetate                                             6β-Hydrocortisol                                                          Hydrocortisone                                                                 11α-Hydrocortione                                                        Hydrocortisone acetate                                                         Hydrocortisone acetate, 3-CMO                                                  Hydrocortisone acetate, 3-CMO:BSA                                              Hydrocortisone 3-CMO                                                           Hydrocortisone 3-CMO:BSA                                                       Hydrocortisone glucuronide                                                     Hydrocortisone hemisuccinate                                                   Hydrocortisone hemisuccinate:BSA                                               3β-Hydroxy-5α-androstan-17-one                                      4-Hydroxyandrostenedione                                                       7α-Hydroxyandrostenedione                                                19-Hydroxy-4-androsten-3,17-dione                                              17β-Hydroxy-4-androsten-3-one                                             16α-Hydroxyandrosterone                                                  3β-Hydroxy-bisnor-5-cholenic acid                                         3β-Hydroxy-5-cholenic acid                                                4β-Hydroxycholesterol                                                     7α-Hydroxycholesterol                                                    7β-Hydroxycholesterol                                                     19-Hydroxycholesterol                                                          20α-Hydroxycholesterol                                                   25-Hydroxycholesterol                                                          Hydroxycholic acid                                                             6β-Hydroxycorticosterone                                                  17-Hydroxycorticosterone                                                       6β-Hydroxycortisol                                                        18-Hydroxy-11-deoxycorticosterone                                              3α-Hydroxy-7,12-dikerocholanic acid                                      18-Hydroxy DOC                                                                 2-Hydroxyestradiol                                                             2-Hydroxyestradiol 17-acetate                                                  2-Hydroxyestradiol 3-methyl ether                                              4-Hydroxyestradiol                                                             6α-Hydroxyestradiol                                                      11α-Hydroxyestradiol                                                     16α-Hydroxy-17β-estradiol                                           16β-Hydroxy-17β-estradiol                                            2-Hydroxyestriol                                                               15α-Hydroxyestriol                                                       2-Hydroxyestrone                                                               4-Hydroxyestrone                                                               16α-Hydroxyestrone                                                       2-Hydroxyestrone-3-methyl ether                                                3β-Hydroxy-5α-etianic acid                                          3β-Hydroxy-5α-etianic acid methyl ester                             3α-Hydroxy-5β-etianic acid                                          3α-Hydroxy-5β-etianic acid methyl ester                             3β-Hydroxy-5β-etianic acid                                           3β-Hydroxy-5β-etianic acid methyl ester                              11β-Hydroxyetiocholanolone                                                16α-Hydroxyetiocholanolone                                               3β-Hydroxy-5-etiocholenic acid                                            3α-Hydroxy-6-ketocholanic acid                                           11α-Hydroxymethyltestosterone                                            6β-Hydroxyprednisolone                                                    17-Hydroxypregnanolone                                                         16α-Hydroxypregnenolone                                                  17-Hydroxypregnenolone                                                         17-Hydroxypregnenolone 3-acetate                                               21-Hydroxypregnenolone                                                         2α-Hydroxyprogesterone                                                   6α-Hydroxyprogesterone                                                   6β-Hydroxyprogesterone                                                    6β-Hydroxyprogesterone acetate                                            6β-Hydroxyprogesterone hemisuccinate                                      6β-Hydroxyprogesterone hemisuccinate:BSA                                  11α-Hydroxyprogesterone                                                  11α-Hydroxyprogesterone acetate                                          11α-Hydroxyprogesterone hemisuccinate                                    11α-Hydroxyprogesterone hemisuccinate:BSA                                11α-Hydroxyprogesterone tosylate                                         11β-Hydroxyprogesterone                                                   12α-Hydroxyprogesterone                                                  16α-Hydroxyprogesterone                                                  17-Hydroxyprogesterone                                                         17-Hydroxyprogesterone 3-CMO                                                   17-Hydroxyprogesterone 3-CMO:BSA                                               18-Hydroxyprogesterone                                                         19-Hydroxyprogesterone                                                         20α-Hydroxyprogesterone                                                  20β-Hydroxyprogesterone                                                   21-Hydroxyprogesterone                                                         6β-Hydroxytestosterone                                                    7α-Hydroxytestosterone                                                   11α-Hydroxytestosterone                                                  11α-Hydroxytestosterone hemisuccinate                                    11α-Hydroxytestosterone hemisuccinate:BSA                                11β-Hydroxytestosterone                                                   16α-Hydroxytestosterone                                                  16β-Hydroxytestosterone                                                   19-Hydroxytestosterone                                                         11α-Hydroxytigogenin                                                     Hyocholic acid                                                                 Hyocholic acid methyl ester                                                    Hyodeoxycholic acid                                                            Hyodeoxycholic acid methyl ester                                               Iodocholesterol                                                                Isoallospirostan-3β,12β-diol                                         Isoandrosterone                                                                14-iso-Equilenin acetate                                                       14-iso-Equilenin methyl ether                                                  Isoergosterone                                                                 Iso-Sarsasapogenin                                                             Kendall's compound "A"                                                         Kendall's compound "B"                                                         Kendall's compound "C"                                                         Kendall's compound "E"                                                         Kendall's compound "E" acetate                                                 Kendall's compound "F"                                                         Kendall's compound "G"                                                         Kendall's compound "H"                                                         Kendall's desoxy compound "B"                                                  11-Ketoandrosterone                                                            Ketocholanic acid                                                              3-Ketocholanic acid                                                            6-Ketocholestanol                                                              6-Ketocholestanol acetate                                                      7-Ketocholestanol                                                              6-Ketocholestenone                                                             7-Ketocholesterol                                                              7-Ketocholesterol acetate                                                      18-Ketocorticosterone                                                          7-Ketodeoxycholic acid                                                         6-Keto-17α-estiadiol                                                     6-Keto-17α-estradiol 6-CMO                                               6-Keto-17α-estradiol 6-CMO:BSA                                           6-Keto-17β-estradiol                                                      6-Keto-17β-estradiol 6-CMO                                                6-Keto-17β-estradiol 6-CMO:BSA                                            16-Keto-17β-estradiol                                                     6-Ketoestriol                                                                  6-Ketoestriol 6-CMO                                                            6-Ketoestriol 6-CMO:BSA                                                        6-Ketoestriol triacetate                                                       6-Ketoestrone                                                                  6-Ketoestrone acetate                                                          6-Ketoestrone 6-CMO                                                            6-Ketoestrone 6-CMO:BSA                                                        16-Ketoestrone                                                                 6-Ketoethynylestradiol                                                         6-Ketoethynylestradiol 6-CMO                                                   6-Ketoethynylestradiol 6-CMO:BSA                                               3-Keto-5α-etianic acid                                                   3-Keto-5β-etianic acid                                                    3-Ketoetiocholanic acid                                                        3-Ketoetiocholanic acid methyl ester                                           11-Ketoetiocholanolone                                                         3-Keto-4-etiocholenic acid                                                     3-Keto-4-etiocholenic acid ethyl ester                                         3-Keto-4-etiocholenic acid methyl ester                                        11-Ketoisoandrosterone                                                         6-Ketolithocholic acid                                                         7-Ketolithocholic acid                                                         12-Ketolithocholic acid                                                        12-Ketolithocholic acid acetate, methyl ester                                  12-Ketolithocholic acid benzoate, methyl ester                                 11-Ketopregnanolone                                                            11-Ketopregnanolone acetate                                                    7-Ketopregnenolone                                                             6-Ketoprogesterone                                                             11-Ketoprogesterone                                                            11-Ketotestosterone                                                            16-Ketotestosrerone                                                            16-Ketotestosterone acetate                                                    11-Kerotigogenin                                                               Lanosterol                                                                     Lithocholic acid                                                               Lithocholic acid acetate                                                       Lithocholic acid acetate methyl ester                                          Medroxyprogesterone                                                            Megestrol acetate                                                              Meprednisone                                                                   Mestranol                                                                      4-Methoxyestradiol                                                             2-Methoxyestrapentol                                                           4-Methoxyestriol                                                               2-Methoxyestrone                                                               4-Methoxyestrone                                                               2-Methoxyestrone 3-methyl ether                                                2-Methoxyethynylestradiol                                                      16α-Methyl-17-acetoprogesterone                                          Methyl-3α-Acetoxycholanate                                               Methyl-7α-acetoxy-3,12-diketocholanate                                   Methyl-3α-acetoxy-12α-hydroxycholanate                             Methyl-3α-acetoxy-12-ketocholanate                                       Methylacetoxylithocholate                                                      Methylandrostanediol                                                           Methylandrostanolone                                                           Methylandrostendiol                                                            Methylchenodeoxycholic diacetate                                               Methylchenodeoxycholate                                                        Methyl cholate                                                                 Methyl cholate 3,7-diacetate                                                   6α-Methylhydrocortisone                                                  Methyldehydrocholate                                                           9(11)-Methyl dehydrotestosterone                                               Methyldeoxycholate                                                             6α-Methyl-11-desoxycortisol                                              17α-Methyldihydrotestosterone 3-CMO                                      Methyldihydrotestosterone                                                      Methyl-3α,12α-dihydroxynorcholanate                                Methyl-3α,12α-diol diacetoxynorcholanate                           6-Methyldiosgenin                                                              6-Methyldiosgenin acetate                                                      6-Methylepoxypregnenolone                                                      7α-Methylestradiol                                                       Methylestradiol 3-methyl ether                                                 1-Methylestrone                                                                7α-Methylestrone                                                         6α-Methyl-17-hydroprogesterone                                           16α-Methyl-17-hydroxyprogesterone                                        Methylhydroxytigogenin                                                         Methylhydroxytigogenin 3-acetate                                               Methylhyodeoxycholate                                                          Methyl lithocholate                                                            7α-Methyl-19-nor-testosterone                                            17α-Methyl-19-nor-testosterone                                           Methyloxyprogesterone                                                          6α-Methylprednisolone                                                    6α-Methylprednisolone acetate                                            6α-Methylprednisolone hemisuccinate                                      6α-Methylprednisolone sodium succinate                                   16β-Methylprednisone                                                      6-Methylpregnenolone                                                           6-Methylpregnenolone acetate                                                   16α-Methylpregnenolone                                                   16β-Methylpregnenolone                                                    16α-Methylprogesterone                                                   16α-Methyl substance "S"                                                 16α-Methyl substance "S" acetate                                         Methyltestosterone                                                             Δ1-Methyltestosterone                                                    17α-Methyl-Δ1-testosterone                                         2-Methyoxyestradiol                                                            2-Methyoxyestradiol 3-methyl ether                                             Methyl-3β,12α-diacetoxycholanate                                    Methyl-3β,12α-diacetoxydeoxycholanate                               Murocholic acid                                                                α-Muricholic acid                                                        β-Muricholic acid                                                         Nandrolone                                                                     Neocholestene                                                                  Nilevar                                                                        6-Nitrocholesteryl acetate                                                     6-Nitrocholesteryl benzoate                                                    2-Nitroestradiol                                                               4-Nitroestrone                                                                 19-Nor-4-androsten-3,17-dione                                                  19-Nor-4-androsten-17α-ethyl-17β-ol-3-one                           19-Nor-4-androsten-17α-ethynyl-17β-ol-3-one                         19-Nor-4-androsten-17β-ol-3-one                                           19-Nor-Androsterone                                                            Nordeoxycholic add                                                             Nordeoxycholic acid diacetate                                                  Nordeoxycholic acid diacetate, methyl ester                                    Nordeoxycholic acid methyl ester                                               Norethandrolone                                                                Norethidrone                                                                   19-Nor-4-ethisterone                                                           Norethynodrel                                                                  Norgestrel                                                                     Norlutin                                                                       19-Norprogesterone                                                             19-Nortestosterone                                                             19-Nortestosterone acetate                                                     19-Nortestosterone benzoate                                                    19-Nortestosterone 3-CMO                                                       19-Nortestosterone dichloroacetate                                             19-Nortestosterone hemisuccinate                                               19-Nortestosterone propionate                                                  19-Nortestosterone sodium sulfate                                              Oxandrolone                                                                    Oxymetholone                                                                   Prednisolone                                                                   Prednisolone acetate                                                           Prednisolone 21-carboxylic acid                                                Prednisolone hemisuccinate                                                     Prednisolone 21-phosphate, disodium salt                                       Prednisone                                                                     Prednisone acetate                                                             Prednisone hemisuccinate                                                       Prednisone 21-hemisuccinate Na salt                                            Pregnanediol                                                                   Pregnanediol diacetate                                                         Pregnanedione                                                                  Pregnanetriol                                                                  Pregnanetriol 3-glucuronide sodium salt                                        Pregnanetriolone                                                               Pregnanolone                                                                   Pregnanolone acetate                                                           Pregnanolone hemisuccinate                                                     4-Pregnen-11β,21-diol-3,20-dione 18-al                                    Pregnenindiol                                                                  Pregnenolone                                                                   Pregnenolone acetate                                                           Pregnenolone acetate oxime                                                     Pregnenolone 20-CMO                                                            Pregnenolone glucuronide                                                       Pregnenolone hemisuccinate                                                     Pregnenolone methyl ether                                                      Pregnenolone sodium sulfate                                                    Pregnenolone tosylate                                                          Δ1-Progesterone                                                          Progesterone                                                                   Progesterone 3-CMO                                                             Progesterone 3-CMO:BSA                                                         Provera                                                                        Reichstein's epi "U"                                                           Reichstein's substance "A"                                                     Reichstein's substance "C"                                                     Reichstein's substance "Dehydro C"                                             Reichstein's substance "D"                                                     Reichstein's substance Epi "E"                                                 Reichstein's substance "E"                                                     Reichstein's substance "Fa"                                                    Reichstein's substance "G"                                                     Reichstein's substance "H"                                                     Reichstein's substance "J"                                                     Reichstein's substance "K"                                                     Reichstein's substance "L"                                                     Reichstein's substance "M"                                                     Reichstein's suhstance "N"                                                     Reichstein's suhstance "O"                                                     Reichstein's substance "P"                                                     Reichstein's substance "Q"                                                     Reichstein's substance "S"                                                     Reichstein's substance "T"                                                     Reichstein's substance "U"                                                     Reichstein's substance "V"                                                     Rockogenin                                                                     Sirsasapogenin                                                                 Sarsasapogenin acetate                                                         β-Sitosterol                                                              β-Sitosterol acetate                                                      Smilagenin                                                                     Smilagenin acetate                                                             Sodium cholate                                                                 Sodium dehydrocholate                                                          Sodium glycochenodeoxycholate                                                  Sodium glycocholate                                                            Sodium glycodehydrocholate                                                     Sodium glycodeoxycholate                                                       Sodium glycohyodeoxycholate                                                    Sodium glycolithocholate                                                       Sodium lithocholate                                                            Sodium taurochenodeoxycholate                                                  Sodium taurocholanic acid                                                      Sodium taurocholate                                                            Sodium taurodehydrocholate                                                     Sodium taurodeoxycholate                                                       Sodium taurohyodeoxycholate                                                    Sodium taurolithocholate                                                       Stanolone                                                                      Stigmastadienone                                                               Srigmasterol                                                                   Stigmasterol acetate                                                           Stilbestrol                                                                    Taurochenodeoxycholic acid                                                     Taurochenodeoxycholic acid sodium salt                                         Taurocholanic acid                                                             Taurocholanic acid sodium salt                                                 Taurocholic acid                                                               Taurocholic acid sodium salt                                                   Taurodehydrocholic acid                                                        Taurodehydrocholic acid sodium salt                                            Taurodeoxycholic acid                                                          Taurodeoxycholic acid sodium salt                                              Taurohyodeoxycholic acid                                                       Taurohyodeoxycholic acid sodium salt                                           Taurolithocholic acid                                                          Taurolithocholic acid sodium salt                                              Testane                                                                        Δ1-Testosterone                                                          Δ1-Testosterone acetate                                                  Δ1-Testosterone benzoate                                                 Δ1-Testosterone 3-CMO                                                    Δ1-Testosterone hemisuccinate                                            Δ1-Testosterone hexahydrobenzoate                                        Δ1-Testosterone propionate                                               Δ1-Testosterone sodium sulfate                                           Testosterone                                                                   Testosterone acetate                                                           Testosterone benzoate                                                          Testosterone 3-CMO                                                             Testosterone 3-CMO:BSA                                                         Testosterone cyclopentylpropionate                                             Testosterone dichloroacetate                                                   Testosterone enol diacetate                                                    Testosterone 3-ethyleneketal                                                   Testosterone glucuronide                                                       Testosterone glucuronide sodium salt                                           Testosterone hemisuccinate                                                     Testosterone hemisuccinate:BSA                                                 Testosterone hexahydrobenzoate                                                 Testosterone phosphoric acid                                                   Testosterone potassium sulfate                                                 Testosterone propionate                                                        Testosterone sodium sulfate                                                    Testosterone tosylate                                                          Tetrahydro compound "A"                                                        Tetrahydro compound "B"                                                        Tetrahydro compound "E"                                                        Tetrahydro compound "E" acetate                                                Tetrahydro compound "F"                                                        Tetrahydrocortexolone                                                          Tetrahydrocorticosterone                                                       Tetrahydrocortisol                                                             Tetrahydrocortisone                                                            Tetrahydrocortisone acetate                                                    Tetrahydro-11-dehydrocorticosterone                                            Tetrahydrodesoxycorticosterone                                                 Tetrahydro-11-desoxycortisol                                                   Tetrahydro DOC                                                                 Tetrahydro hydrocortisone                                                      Tetrahydro substance "Q"                                                       Tetrahydro substance "S"                                                       Tetrahydro substance "S" 21-acetate                                            TH "A"                                                                         TH "B"                                                                         TH "E"                                                                         TH "F"                                                                         TH "S"                                                                         TH "S" 21-acetate                                                              Theelol                                                                        Tigogenin                                                                      Tigogenin acetate                                                              Transdehydroandrosterone                                                       Transestriol                                                                   Triamcinolone                                                                  Triamcinolone acetonide                                                        Triamcinolone diacetate                                                        Triendiol                                                                      Triketocholanic acid                                                           Urocortisol                                                                    Urocortisone                                                                   Ursocholanic acid                                                              Ursodeoxycholic acid                                                           Wintersteiner's compound "A"                                                   Wintersteiner's compound "B"                                                   Wintersteiner's compound "D"                                                   Wintersteiner's compound "F"                                                   Wintersteiner's compound "G"                                                   ______________________________________                                    

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Although preferred embodiments of the invention are described herein in detail, it will be understood by those skilled in the art that variations may be made thereto without departing from the spirit of the invention or the scope of the appended claims. 

What is claimed is:
 1. An in vivo method for imaging breast and ovarian cancers in non-prostatic tissue of a patient comprising:injecting a patient with a steroid selected from the group consisting of glucocorticoids, mineralcorticoids, androgens, antiestrogens and progestin, which induces the cancer cells to produce prostate specific antigen, said cancer cells having receptors for the injected steroid, injecting a patient with antibodies which bind to prostate specific antigen produced by non-prostatic tissue of the patient, said antibodies being labeled with imaging agents; allowing said antibodies to incubate in vivo and bind prostate specific antigen associated with a cancer; and detecting presence of said imaging agents of bound antibodies localized to the cancer.
 2. A method of claim 1, wherein the antibodies are selected from the group consisting of: polyclonal antibodies, monoclonal antibodies, antibody fragments, antibody constructs, single chain antibodies and bifunctional antibodies.
 3. The method of claim 1, wherein the antibodies are labeled with a radioisotope and the patient is imaged with a photoscanning device.
 4. A method of claim 3, wherein the radioisotope is selected from the group consisting of: ²²⁷ Ac, ²¹¹ AT, ¹²⁸ BA, ¹³¹ Ba, ⁷ Be, ²⁰⁴ Bi, ²⁰⁵ Bi, ²⁰⁶ Bi, ⁷⁶ Br, ⁷⁷ Br, ⁸² Br¹⁰⁹ Cd, ⁴⁷ CA, ¹¹ C, ¹⁴ C, ³⁶ Cl, ⁴⁸ Cr, ⁵¹ Cr, ⁶² Cu, ⁶⁴ Cu, ⁶⁷ Cu, ¹⁶⁵ Dy, ¹⁵⁵ Eu, ¹⁸ F, ¹⁵³ Gd, ⁶⁶ Ga, ⁶⁷ Ga, ⁶⁸ Ga, ⁷² Ga, ¹⁹⁸ Au, ³ H, ¹⁶⁶ Ho, ¹¹¹ In, ^(113m) In, ^(115m) In, ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸⁹ Ir, ^(191m) Ir, ¹⁹² Ir, ¹⁹⁴ Ir, ⁵² Fe, ⁵⁵ Fe, ⁵⁹ Fe, ¹⁷⁷ Lu, ¹⁵ O, ^(191m-191) OS, ¹⁰⁹ Pd, ³² P, ³³ P, ⁴² K, ²²⁶ Ra, ¹⁸⁶ Re, ¹⁸⁸ Re, ^(82m) Rb, ¹⁵³ Sm, ⁴⁶ Sc, ⁴⁷ Sc, ⁷² Se, ⁷⁵ Se, ¹⁰⁵ Ag, ²² Na, ²⁴ Na, ⁸⁹ Sr, ³⁵ S, ³⁸ S, ¹⁷⁷ Ta, ⁹⁶ Tc, ^(99m) Tc, ²⁰¹ Tl, ²⁰² Tl, ¹¹³ Sn, ^(117m) Sn, ¹²¹ Sn, ¹⁶⁶ Yb, ¹⁶⁹ Yb, ¹⁷⁵ Yb, ⁸⁸ Y, ⁹⁰ Y, ⁶² Zn, and ⁶⁵ Zn.
 5. A method of claim 4, wherein the radioisotope is selected from the group consisting of: ¹³¹ I, ¹²⁵ I, ¹¹¹ In, ^(99m) Tc, ⁹⁰ Y, ¹⁸⁶ Re, ¹⁵³ Sm, ⁶⁷ Ga, ²⁰¹ Tl, ⁷⁷ Br, and ¹⁸ F.
 6. The method of claim 1, wherein the antibodies are labeled with a metal attached covalently to create a paramagnetic conjugate and the patient is imaged by magnetic resonance imaging.
 7. A method of claim 6, wherein the metal is selected from the group consisting of: gadolinium, terbium, tin, iron and isotopes thereof. 